Fig 2: Analysis of miRNA-seq. (A) Length distribution of miRNAs in the six libraries. (B) Volcano plot displaying miRNA expression alterations in HIF1A-KO cells compared to those of the control. (C) Top 20 KEGG pathways enrichment of DEMs. (D) GO analysis of DEMs through diverse GO categories.

Fig 3: MTA1/HIF1A/AQP4 promotes the apoptosis of colon epithelial cells.A TUNEL staining of the cell apoptosis of colon tissue in DSS-induced experimental colitis. B The cell viability rate detected by CCK-8 assay in LPS-treated cells with MTA1 silencing. C The apoptotic rate detected by flow cytometry in LPS-treated cells with MTA1 silencing. D The cell viability rate detected by CCK-8 assay after AQP4 was overexpressed in LPS-treated cells with MTA1 silencing. E The apoptotic rate detected by flow cytometry after AQP4 was overexpressed in LPS-treated cells with MTA1 silencing. *p < 0.05; **p < 0.01; ***p < 0.001. The measurement data were expressed as mean ± standard deviation. Data between two groups were compared by the unpaired student’s t test, and data among multiple groups were compared by one-way ANOVA, followed by Tukey’s post hoc tests. The experiment was conducted three times independently.

Fig 4: CCL5 derived from CAFs upregulates HIF1α in liver cancer cells.A The expression of HIF1α in Huh7 and Hep3B cells treated with hCCL5 (20 ng/ml and 100 ng/ml) was detected by RT-PCR and western blotting. B The protein expression of HIF1α in Huh7 and Hep3B cells treated with the CM of CAFs, a CCL5 neutralizing antibody or CCR3/CCR5 antagonists was assessed using western blotting. C HIF1α expression of Huh7 and Hep3B cells were tested by immunofluorescence staining and photographed under a confocal microscopy when treated with CAF-CM or CAF-CM treated with a CCL5 neutralizing antibody. Scale bar: 20 μm. D Immunofluorescence staining of HIF1α protein was detected in four groups of tumor samples including Huh7 cells, Huh7 cells co-injected with CAFs, Huh7 cells co-injected with shCCL5 CAFs and shCCR3/5 Huh7 cells co-injected with CAFs. Scale bar: 100 μm. E Pearson correlation analysis showed that HIF1A was positively correlated with the expression of CCL5 in 66 liver cancer samples (r = 0.6312, p < 0.0001). N = 5 per group. Mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, compared with the tumor control group.

Fig 5: Activation of HIF1A signaling in early prostatic lesions and its pharmacological targeting.(A) UMAP of unsupervised clustering of prostatic cells from PtenL2/L2 and Pten(i)pe-/- mice, 3 months AGI, and (B) dot plot depicting cell lineage–specific markers. (C) KEGG pathway analysis of genes up-regulated in Pten-null prostatic luminal cells. (D) Violin plots depicting the levels of HIF1A signaling–related transcripts in luminal-A and -C cells. ***P < 0.001, Wilcoxon rank sum test. (E) Representative immunohistochemical (IHC) staining of HIF1A and carbonic anhydrase 9 (CA-IX) in sections of dorsolateral prostate (DLP) of PtenL2/L2 and Pten(i)pe-/- mice 3 months AGI. n = 3 mice per condition. Scale bars, 100 µm. (F) Pimonidazole IHC staining in the DLP of PtenL2/L2 and Pten(i)pe-/- mice 3 months AGI. n = 3 mice per condition. Scale bars, 100 µm. Prostate weight (G) and representative H&E staining (H, left) and cleaved caspase 3 (C3) IHC staining (H, right) of Pten(i)pe-/- mice, 3 months AGI, treated for 5 days with vehicle (n = 3) or PX-478 (n = 6). *P < 0.05, two-tailed t test. Scale bars, 100 µm. Prostate weight (I) and H&E-stained DLP (J) of PtenL2/L2 (n = 4 per condition) and Pten(i)pe-/- (n = 6 per condition) mice (10 months AGI) treated for 28 days with vehicle or PX-478. n.s., not significant, P = 0.05; **P < 0.01, two-tailed t test. Scale bars, 100 µm. (K) Quantification of Ki-67–positive epithelial cells in the DLP of vehicle- (n = 3) and PX-478–treated (n = 4) Pten(i)pe-/- mice, 10 months AGI. Five to 10 images per prostatic section were analyzed. *P < 0.05, two-tailed t test.